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12 July 2018 : Laboratory Research  

Genistein and Myd88 Activate Autophagy in High Glucose-Induced Renal Podocytes In Vitro

Yuanyuan Wang1ABCDEF, Ying Li1AE*, Tao Zhang1BCD, Yanqing Chi1BCD, Maodong Liu1BCD, Ying Liu2CF

DOI: 10.12659/MSM.910868

Med Sci Monit 2018; 24: LBR4823-4831

Abstract

BACKGROUND: Renal podocyte damage plays a crucial role in the development of diabetic nephropathy. Genistein is derived from a leguminous plant, and MyD88 and TRIF are adaptor molecules in the Toll-like receptor (TLR) signaling pathway, which may play a role in autophagy. In this study, we utilized an in vitro high glucose (HG)-treated podocyte model to investigate the effects and underlying mechanisms of Genistein and MyD88 or TRIF siRNA induced autophagy and renal protection.

MATERIAL AND METHODS: An immortalized mouse podocyte cell line was treated with HG, Genistein, chloroquine, and/or transfected with specific Myd88 and TRIF siRNAs. The formation of autophagosomes and related autophagic vacuoles were monitored by transmission electron microscopy. The expression of autophagy-related factors and podocyte structure and functional markers, including LC3, p62, p-mTOR, synaptopodin, and nephrin, were measured by Western blot, and LC3 and p-mTOR expression were also assessed by immunofluorescence.

RESULTS: We showed that HG transiently (after 6-h exposure) induced expression of the autophagy activation marker LC3-II in podocytes. Genistein treatment induced autophagy in both normal and HG-treated podocytes through inactivating mTOR signaling. Moreover, Genistein protected podocytes against chloroquine in HG-cultured conditions in vitro by maintaining the level of autophagy-related proteins. In addition, MyD88 siRNA downregulated expression of autophagy-related proteins, whereas Genistein treatment reversed these effects.

CONCLUSIONS: This study demonstrated that Genistein-induced autophagy could be a potential treatment strategy for glomerular diseases.

Keywords: Genistein, Podocytes

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750